title: Bradykinin controls pool size of sensory neurons expressing functional δ-opioid receptors.
abstract: Analgesics targeting the δ-opioid receptor (DOR) may lead to fewer side effects than conventional opioid drugs, which mainly act on μ-opioid receptors (MOR), because of the less abundant expression of DOR in the CNS compared with MOR. Analgesic potential of DOR agonists increases after inflammation, an effect that may be mediated by DOR expressed in the peripheral sensory fibers. However, the expression of functional DOR at the plasma membrane of sensory neurons is controversial. Here we have used patch-clamp recordings and total internal reflection fluorescence microscopy to study the functional expression of DOR in sensory neurons from rat trigeminal (TG) and dorsal root ganglia (DRG). Real-time total internal reflection fluorescence microscopy revealed that treatment of TG and DRG cultures with the inflammatory mediator bradykinin (BK) caused robust trafficking of heterologously expressed GFP-tagged DOR to the plasma membrane. By contrast, treatment of neurons with the DOR agonist [d-Ala(2), d-Leu(5)]-enkephalin (DADLE) caused a decrease in the membrane abundance of DOR, suggesting internalization of the receptor after agonist binding. Patch-clamp experiments revealed that DADLE inhibited voltage-gated Ca(2+) channels (VGCCs) in 23% of small-diameter TG neurons. Pretreatment with BK resulted in more than twice as many DADLE responsive neurons (54%) but did not affect the efficacy of VGCC inhibition by DADLE. Our data suggest that inflammatory mediator-induced membrane insertion of DOR into the plasma membrane of peripheral sensory neurons may underlie increased DOR analgesia in inflamed tissue. Furthermore, the majority of BK-responsive TG neurons may have a potential to become responsive to DOR ligands in inflammatory conditions.
authors.affiliation: School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, LS2 9JT Leeds, United Kingdom.
authors.forename: Louisa
authors.initials: L
authors.lastname: Pettinger
authors.forename: Sylvain
authors.initials: S
authors.lastname: Gigout
authors.forename: John E
authors.initials: JE
authors.lastname: Linley
authors.forename: Nikita
authors.initials: N
authors.lastname: Gamper
mesh_terms: D000818:Animals; D001920:Bradykinin / Q000494:pharmacology*; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism; D015220:Calcium Channels / Q000502:physiology; D002211:Capsaicin / Q000494:pharmacology; D002452:Cell Count; D002462:Cell Membrane / Q000378:metabolism; D016308:Enkephalin, Leucine-2-Alanine / Q000494:pharmacology; D005260:Female; D015640:Ion Channel Gating / Q000502:physiology; D008297:Male; D008856:Microscopy, Fluorescence; D018408:Patch-Clamp Techniques; D051381:Rats; D017208:Rats, Wistar; D017465:Receptors, Opioid, delta / Q000187:drug effects / Q000378:metabolism*; D011984:Sensory Receptor Cells / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology*; D050916:TRPV Cation Channels / Q000502:physiology; D012668:Trigeminal Ganglion / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
chemical_list: D015220:Calcium Channels; D017465:Receptors, Opioid, delta; D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat; D016308:Enkephalin, Leucine-2-Alanine; D015740:Calcitonin Gene-Related Peptide; D002211:Capsaicin; D001920:Bradykinin